Objective To investigate the radiosensitivity of stem cells in pancreatic adenocarcinoma PANC‐1 cell line and the possible mechanism of its radiation resistance .Methods Using flow cytometry ,the cells were isolated and sorted into CD44+CD24+ ,CD44-CD24+ ,CD44+CD24- ,and CD44-CD24- .Multi‐target click model was used to fit cell survival curves and deter‐mine the sensitizer enhancement ratio .The apoptosis and cycle distribution of the four cell subsets were determined using flow cy‐tometry ,and the level of ROS was determined by DCFH‐DA probe .Results The ratio of CD44+ and CD24+ in the sorted PANC‐1 cell line was 92 .0% and 4 .7% respectively .Before radiation ,there was no statistically significant difference between four groups (P>0 .05);After treated with 6MV‐X ray ,The ratio of apoptosis was the lowest in CD44+CD24+ (P<0 .01);The percentage G0/G1 cell was the highest in CD44+CD24+ (P<0 .01) ,the sensitizer enhancement ratio of CD44+ CD24- ,CD44-CD24+ and CD44-CD24- were 1 .61 ,1 .81 ,1 .94 ,respectively .The level of ROS in CD44+CD24+ was lower (P<0 .01) .Conclusion Tumor stem cells of pancreatic adenocarcinoma have properties of a lower level of ROS and relative stationary that maybe the reasons of radio resist‐ance .

Objective To compare the dissolution behavior between domestic trepibutone tablets and original reference product, and provide a basis for evaluating the quality consistency of generic drugs. Methods Four dissolution media recommended by Japanese Orange Book and a domestic standard dissolution media were selected to determined the dissolution profile,and f2 factor was calculated to investigate the consistency of stripping curves. Results In water,pH 4.0 and pH 1.2,the f2 of domestic formulation and reference formulation was under 50,and the dissolution profile was inconsistent.Dissolution behavior of domestic preparations of different manufacturers was dissimilar.In water,the f2 of domestic preparations of different batches of the same manufacturer was over 99.9,and the dissolution behavior was similar. Conclusion The dissolution method of existing domestic standard can not distinguish the dissolution behavior of different products,and it should be revised and completed.There is still great difference in quality between the domestic preparations and reference preparations.

Objective To examine the effect of modified electroconvulsive therapy (MECT) on the plasma oxida-tive stress level in bipolar depression. Methods Forty-two patients with bipolar depression were randomly divided into two groups. The intervention group (n=18) received antidepressants and 12 times MECT for 6 weeks and the control group (n=24) received antidepressants and Li2CO3 for 6 weeks. The Chinese versions of the 17 items Hamilton Depression Rating Scale (HAMD-17), Young Mania Rating Scale (YMRS), Clinical Global Impression Scale (CGI-S) and Treatment Emergent Symptom Scale (TESS) were used to assess participants at baseline and after 6 weeks of treatment. The plasma levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA)were detected at baseline and after 6 weeks to assess the level of oxidative stress. Results Repeated measures analysis of variance showed that the plasma level of SOD was higher in MECT group than in Li2CO3 group (F=15.26, P<0.01), and the level of MDA was higher in Li2CO3 group (F=18.18, P<0.01). The interactive effect of group and time was significant in GSH-Px level (F=6.39, P=0.02). The level of GSH-Px was lower in MECT group than in Li2CO3 group after 6 weeks (P<0.05). The CAT level was higher in the response patients than in non-response patients after 6 weeks (P<0.05). Con-clusions Both MECT treatment and Li2CO3 treatment can alter oxidative stress levels in patients with bipolar depression. The mechanisms underlying its therapeutic regimen may correct the imbalance of the plasma CAT level.

Objective To investigate the diagnosis and surgical treatment of intracaval venous tumors. Methods Clinical data of 6 cases were retrospectively analyzed, including signs and symptoms diagnostic means such as type-B ultrasound, CTA, MRA, surgical procedures and prognosis. Results All six cases received type-B ultrasonic examination, final definite diagnosis was achieved by CTA exam in 2 cases and through MRA in 4 cases. Heart involvement was found in 3 cases. All patients underwent a surgery. According to the extent of the tumor,3 cases had thoraco-abdominal incision,3 cases with extracorporeal circulation and right atrium opening. All of the tumors were completely resected. Pathological exam revealed that 4 cases were of leiomyomatosis and 2 cases were of leiomyosarcoma. One case with leiomyosarcoma died of liver disfunction postoperatively.The other 5 cases recovered without major complications. An average 51 months of follow-up found no recurrence. Conclusions CT and MRI are the mainstay for the diagnosis,and MRI can provide clear anatomy image to the surgeons, help choose the surgical procedures. The one-stage operation is effective. During the operation, the main branches of the vena cava system should be detected, and the attachment of the tumor should be found and removed thoroughly to prevent the recurrence of the tumor. When the attachment point is lower than the iliac vein level, ligation of the involved iliac vein should be mandatory.

Objective To investigate the effect of signal transducer and activator of transcription-3(STAT-3)-mediated non-dependent vascular endothelial growth factor receptor-2(VEGFR-2) pathway on the radiosensitivity of non-small cell lung cancer (NSCLC) cells and its possible mechanism.Methods NSCLC cells were divided into control group, apatinib (VEGFR-2 inhibitor) group, apatinib+S3I-201(STAT-3 inhibitor) group, radiation (RT) group, RT+apatinib group, and RT+apatinib+S3I-201 group.Q-PCR and Western blot were used to measure the mRNA and protein expression of VEGFR-2, STAT-3, other proteins involved in the signaling pathway, hypoxia-inducible factor 1α(HIF-1α), and cyclin D1.The cellular radiosensitivity was determined by colony-forming assay and cell apoptosis and cell cycle distribution were evaluated by flow cytometry.Results For Calu-1 cells, there was no significant difference in the expression of VEGFR-2 mRNA and STAT-3 mRNA between the apatinib group and the control group (P>0.05);the apatinib group had significantly lower expression of HIF-1α mRNA and cyclin D1 mRNA than the control group (P<0.05);the apatinib+S3I-201 group had significantly lower mRNA and phosphorylated protein expression of VEGFR-2, STAT-3, and related downstream target proteins than the control group (F=304.54, P<0.01;F=118.99, P<0.01, F=144.34, P<0.01;F=529.66, P<0.01);compared with the control group, the apatinib+S3I-201 group and the apatinib group showed increases in apoptosis rate and proportion of cells in G2+M phase, and the apatinib+S3I-201 group had significantly greater increases than the apatinib group (F=72.37, P<0.01).Compared with Calu-1 cells, the radiosensitizing effect of apatinib on A549 cells was limited (SER[sensitizer enhancement ratio]=1.39).The radiosensitizing effect of apatinib+S3I-201 on A549 cells was significantly higher than that of apatinib (SER:1.72 vs.1.39, P=0.000).Conclusions The inhibition of STAT-3 can enhance the radiosensitivity of NSCLC cells.When VEGFR-2 is inhibited by apatinib, activated STAT-3 regulates the expression of cyclin D1 directly or indirectly to affect the radiosensitivity of NSCLC cells.The inhibition of VEGFR-2 and STAT-3 has a good radiosensitizing effect on NSCLC cells.

The general characteristics, outcomes and risk factors of the patients with aortic dissection (AD) were evaluated in a single medical center. From January 2002 to December 2008, 284 patients with AD were treated and followed-up at our institution, including 105 cases of type A AD and 179 cases of type B AD. The patients in each type were divided into three groups according to management: medical treatment group (A or B), open surgery group (A or B), and stent-graft group (A or B). The characteristics and follow-up outcomes were compared between the groups or subgroups. The results showed that there was significant difference in the prognosis for type A AD between medical treatment group and open surgery group, but there was no significant difference in the prognosis for type B AD between medical treatment group and stent-graft group. Independent risk factors of follow-up mortality for patients with type A AD included a history of atherosclerosis (HR, 3.807; 95% confidence interval [CI], 1.489 to 7.611; P=0.003), in-hospital hypotension/shock (HR, 4.687; 95% CI, 1.846 to 11.900; P=0.001), in-hospital myocardial ischemia or infarction (HR, 3.734; 95% CI, 1.613 to 8.643; P=0.002), pleural effusion (HR, 2.210; 95% CI, 1.080 to 4.521; P=0.030), branch vessel involvement (HR, 2.747; 95% CI, 1.202 to 6.278; P=0.016) and surgical treatment (HR, 0.177; 95% CI, 0.063 to 0.502; P=0.001). And there were insignificant independent predictors for mortality of the patients with type B AD. It was concluded that there were significant differences in characteristics and one year mortality between type A AD and type B AD, but after one year, there was no significant difference in the mortality and complications of them. There were several discordant risk factors of AD, such as female gender, age, thrombus, abrupt onset of pain that were considered as the risk factors in some papers. And there was no definite risk factor of mortality in this study in the patients with type B AD.

OBJECTIVETo prepare freeze-dried long-circulation oridonin liposomes with optimized parameters.

METHODSEthanol injection method followed by freeze-drying was used to prepare the liposomes. Sephadex column was used to purify liposomes. Effects of formulation factors on entrapment efficiency of long-circulation oridonin liposomes were studied. The particle size, distribution and in vitro release were determined. Pharmacokinetics of oridonin liposomes in rats was determined by HPLC and the pharmacokinetic parameters calculated by Kinetica(TM) software were compared with conventional oridonin liposomes and solution.

RESULTSThe optimized lipid formulation for long-circulation liposomes was composed of soy lecithin, cholesterol and DSPE-PEG 2000 with a ratio of 1:0.5:1.8(w/w). The ratio of drug to lipid was 1:6. Freeze-drying protectant was a mixture of glucose and mannitol (3:1). The entrapment efficiency (EE) of long-circulation oridonin liposomes was about 65%. The particle size of liposomes after hydrolyzation was 164 nm with good DPI. The liposomes showed a sustained drug release in vitro. Intravenous injected oridonin fitted with two-compartment pharmacokinetic model. The MRT of long-circulation liposomes was 2 times and 6 times and AUC was about 2 times and 3 times of conventional liposomes and oridonin solution, respectively.

CONCLUSIONFreeze-dried liposomes with high EE have been obtained by the proposed approach. This long-circulation liposomes extend oridonin half time and significantly increase AUC in rats.

Animals ; Delayed-Action Preparations ; Diterpenes, Kaurane ; administration & dosage ; pharmacokinetics ; Drug Stability ; Freeze Drying ; Liposomes ; administration & dosage ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution

Objective To determine the effects of endostatin on the expression of vascular endothelial growth factor receptor?2 ( VEGFR?2 ) in non?small cell lung cancer cells ( human A549 lung adenocarcinoma cells and human Calu?1 lung carcinoma cells) , and to investigate the possible mechanisms underlying its radiosensitizing effect. Methods The CCK8 method was used to determine the inhibitory effect of endostatin on cell proliferation and calculate the drug concentration that caused a 20% reduction in cell proliferation within 24 h ( IC20 ) . RT?PCR and Western blot assays were used to assess the mRNA and protein expression of VEGFR?2, proteins within its related signaling pathways, and HIF?1α, respectively. The radiosensitivity of cells in each group was determined by colony formation assay;cell apoptosis and cell cycle distribution were determined by flow cytometry. Comparison of mean values between multiple samples was made by one?way analysis of variance, and comparison of mean values between two samples was made by t test. Results Endostatin significantly inhibited the proliferation of Calu?1 cells ( F=50?36,P<0?01) with an IC20 of 296?5 μg/ml;the mRNA and protein expression of VEGFR?2 and HIF?1α was also significantly inhibited in endostatin?treated Calu?1 cells ( F=25?43,10?44, all P<0?05) . Moreover, the phosphorylation of Akt, ERK 1/2, and p38 was significantly reduced in endostatin?treated Calu?1 cells ( F=2?89,0?24, 1?09, all P<0?05) . The radiosensitivity enhancement ratios for Calu?1 cells and A549 cells were 1?38 and 1?09, respectively. Endostatin significantly induced apoptosis ( F=44?15, P<0?01) and G2/M blockage ( F= 104?24, P< 0?01 ) in Calu?1 cells. Conclusions Endostatin induces apoptosis and enhances radiosensitivity in Calu?1 cells with high expression of VEGFR?2, but it has a limited impact on A549 cells with low expression of VEGFR?2.

BACKGROUND:Human mesenchymal stem cells derived from Wharton's jelly(WJCs)display the characteristics of MSCs as defined by the International Society for Cellular Therapy.They can be differentiated into bone,cartilage,adipose,muscle,and neural cells.They can also support the expansion of other stem cells,be weli-tolerated by the immune system,and have the ability to home to tumors.OBJECTIVE:To investigate biological changes of WJCs and brain tumor stem cells(BTSCs)co-cultured in vitro.METHODS:WJCs cultured by situ cultivation and BTSCs used enzyme digestion way respectively,and gathering the 3rd passage of WJCs though subculturing as well as BTSCs.Two kinds of cells co-cultured in 24-well plates in serum-free medium (SFM)without any growth factor.3 and 7 days after co-cultured respectively,CD133 expression of suspension cells in the 24-well plates were identified by flow cytometry,and immunofluorescence was performed for Nestin and glial fibrillary acidic protein (GFAP)expression of adherent cells.Co-culture supernatant(CCS)re-suspended 3~(rd) passage of BTSCs and cultured into 96-well plates at day 3,which were used to determine the difference in cell growth curve in both groups using a microplate reader.RESULTS AND CONCLUSION:With the cocultivation days increasing,the phenomenon that tumor sphere cells began to be decomposed,adherent and differentiated observed by an inverted microscope.BTSCs in the co-cultured group expressed GFAP and Nestin when adherent and differentiated.The higher degree of malignant brain tumor tissue used in culturing BTSCs was,the higher expression of CD133 in BTSCs was.CD 133~+ in BTSCs declined when co-cultured with WJCs.Growth curve of brain tumor stem cells cultured in CCS compared with in SFM at day 3,which indicates that the proliferation of BTSCs inhibited obviously.Results indicated that CD 133~+ expression and proliferative capacity of BTSCs went down and BTSCs underwent differentiation during the co-culture in vitro.

BACKGROUND:Bone marrow mesenchymal stem cells have low immunogenicity and immunomodulatory effect,but there are seldom reports concerning the immunomodulatory effect of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord and its mechanims.OBJECTIVE:To investigate the immunomodulatory effects and mechanisms of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord on varient peripheral blood T lymphocytes.METHODS:Mesenchymal stem cells were isolateded from Wharton's jelly of human umbilical cord by tissue culture.T lymphocytes from human peripheral blood were stimulated by phytohemagglutinin and co-cultured with umbilical cord Wharton's jelly-derived mesenchymal stem cells and umbilical cord Wharton's jelly-derived mesenchymal stem cells supernatant respectively to measure A value following 72 hours of coculture using multifunctional microplate reader.Expression of cytokines including transforming growth factor-beta 1(TGF-β1)and interferon-y(IFN-γ)was evaluated by enzyme-labeled immunosorbent assay.RESULTS AND CONCLUSION:Wharton's jelly-derived mesenchymal stem cells could inhibite the proliferation of T lymphocytes induced by phytohemagglutinin.The proliferation inhibition rate was 56%(P＜0.01).Wharton's jelly-derived mesenchymal stem cells supernatant also had inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin,in a dose-dependent fashion.The proliferation inhibition rates were 8.3% and 27% respectively in the 50% Wharton's jelly-derived mesenchymal stem cells supernatant and 100% Wharton's jelly-derived mesenchymal stem cells supematant groups(P＜0.05).Wharton's jelly-derived mesenchymal stem cells significantly decreased γ-interferon secrted from T-lymphocytes(P＜0.05).The secretion of TGF-β1 was lower in the coculture of Wharton's jelly-derived mesenchymal stem cells and T lymphocytes group than Wharton's jelly-derived mesenchymal stem cells alone group(P＜0.05).These indicated that Wharton's jelly-derived mesenchymal stem cells and Wharton's jelly-derived mesenchymal stem cells supernatant have inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin.The mechanims may be associated with cell contant and inhibition of v-interferon secrted from T-lymphocytes.